Species Identification and Antibiotic Susceptibility of Clinical Isolates of Enterococci Isolated from Hospitalized Patients and Outpatients in Split and Dalmatia County, Croatia

Zvonimir Barišić 1, Sanda Sardelić 2, Vinko Zoranić 1, Ivanka Batarilo 1, Merica Carev 1, Volga Punda-Polić 2

1 Public Health Institute, Split, Croatia, 2 University Hospital & Medical School, University of Split, Split, Croatia

Abstract: Identification up to species level and testing antibiotic susceptibility in 235 consecutive clinical isolates of enterococci collected from hospitalized patients and outpatients in Split and Dalmatia County, Croatia during first half of 1997.
E. faecalis was identified in 212 isolates and E. faecium in 23 isolates.
Ampicillin-resistant were 10,64% isolates. There were no beta-lactamase producing isolates. High-level resistant (HLR) gentamicin isolates were 28,94% strains and 45,53% had HLR to streptomycin. Ciprofloxacin-resistant and intermediate were 63,4% isolates. There were no vancomycin-resistant isolates. Nitrofurantoin-resistant were 2,82% of urine isolates.
Statisticaly significant differences in enterococcal resistance among hospitalized and outpatients have been noted in resistance to ampicillin, gentamicin and streptomicin, but difference have not been significant for vancomycin, ciprofloxacin and nitrofurantoin.

Introduction: Enterococci are important causes of community acquired and nosocomial infections. The increased use of antibiotics, especially in hospitals, selects strains of enterococci which are multiple resistant to antibiotics.
The objectives of this research are to establish what species of enterococci prevail as cause of infection in population of Split and Dalmatia County, and to find out if there is a difference in species distribution and frequency of enterococci isolated from clinical samples of hospitalized patients and outpatients, as well as their susceptibility to the most used antibiotics in treatment of enterococcal infections: ampicillin, gentamicin, streptomycin, vancomycin, ciprofloxacin, and, only in cause of urinary tract infection, to nitrofurantoin.

Methods: In this study 235 isolates of enterococci from urine, blood, sperm, wound, cervix, ear and vagina of hospitalized patients and of outpatients in the territory of Split and Dalmatia County, Croatia during first six months of 1997 were tested.
Identification up to the species level was made by Falcam and Collins method and by API 20 Strep (bioMerieux, France).
Antibiotic susceptibility to ampicillin was made by microbroth dilution, to gentamicin and streptomycin by agar dilution screening method, and to vancomycin by E test (AB Biodisk, Sweden). Disk diffusion method was used for testing susceptibility to ciprofloxacin and, in case of urine isolates, to nitrofurantoin. Chi-squared test was used to test difference in resistance among hospital and community isolates of enterococci.

Results: From 235 clinical isolates of enterococci, 106 were from  outpatients (104 E. faecalis and 2 E. faecium) and 129 were from hospitalized patients (108 E. faecalis and 21 E. faecium).
Ampicillin-resistant were 10,64% isolates. There were no beta-lactamase producing isolates. High-level resistant (HLR) gentamicin isolates were 28,94% strains and 45,53% had HLR to streptomycin. Ciprofloxacin-resistant and intermediate were 63,4% isolates. There were no vancomycin-resistant isolates. Nitrofurantoin-resistant were 2,82% of urine isolates.
Ampicillin-resistant were 17,1% isolates from hospitalized patients and 2,8% from outpatients. High-level resistant (HLR) gentamicin isolates were 43,4% isolates from hospitalized patients and 11,3% from outpatients, and HLR streptomycin isolates were 56,6% and 32,1%, respectively. Ciprofloxacin-resistant and intermediate were 68,2% hospitalized patients isolates and 57,6% outpatients isolates. There were no vancomycin-resistant isolates. Nitrofurantoin-resistant were 5,9% of hospitalized patients urine isolates and there were no resistant isolates from outpatients.
Statisticaly significant differences in enterococcal resistance among hospitalized and outpatients have been noted in resistance to ampicillin, gentamicin and streptomicin, but difference have not been significant for vancomycin, ciprofloxacin and nitrofurantoin.

Conclusion: E. faecalis was the species that was mostly isolated from clinical samples.
E. faecium was more frequently isolated from samples collected from hospitalized patients. Statisticaly significant difference in resistance among hospitalized and outpatients was noted in resistance to most clinically used antibiotics: ampicillin, gentamicin and streptomicin, but difference was not significant for vancomycin (there were no vancomycin-resistant isolates at all), ciprofloxacin (it is not antibiotic of first choice to treat enterococcal infections), and nitrofurantoin (antibiotic most frequently used for treating  urinary tract infections in outpatients).